[go: up one dir, main page]

ES2609919T3 - Enzymes conjugated to antibodies using a heterobifunctional PEG linker - Google Patents

Enzymes conjugated to antibodies using a heterobifunctional PEG linker Download PDF

Info

Publication number
ES2609919T3
ES2609919T3 ES06758689.1T ES06758689T ES2609919T3 ES 2609919 T3 ES2609919 T3 ES 2609919T3 ES 06758689 T ES06758689 T ES 06758689T ES 2609919 T3 ES2609919 T3 ES 2609919T3
Authority
ES
Spain
Prior art keywords
conjugate
scaffolding
antibody
staining
generation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
ES06758689.1T
Other languages
Spanish (es)
Inventor
Christopher Bieniarz
Jennifer Wong
Mark Lefever
Jerome W. Kosmeder
Julia Ashworth-Sharpe
Casey A. Kernag
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ventana Medical Systems Inc
Original Assignee
Ventana Medical Systems Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ventana Medical Systems Inc filed Critical Ventana Medical Systems Inc
Application granted granted Critical
Publication of ES2609919T3 publication Critical patent/ES2609919T3/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
    • A61K47/6815—Enzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/44—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Conjugado de anticuerpo-resto generador de señal que comprende un anticuerpo unido covalentemente a un resto generador de señal mediante un conector de PEG heterobifuncional, donde el conjugado tiene la fórmula general:**Fórmula** donde Ab es un anticuerpo, SM es un resto generador de señal que lleva un enzima o un punto cuántico, n >= 4 a 12 y s >= 1 hasta 10.Conjugate of antibody-signal generating moiety comprising an antibody covalently linked to a signal generating moiety via a heterobifunctional PEG linker, where the conjugate has the general formula: ** Formula ** where Ab is an antibody, SM is a moiety signal generator that carries an enzyme or a quantum dot, n> = 4 to 12 and s> = 1 to 10.

Description

imagen1image 1

imagen2image2

imagen3image3

imagen4image4

imagen5image5

imagen6image6

imagen7image7

imagen8image8

imagen9image9

imagen10image10

imagen11image11

imagen12image12

5 5

15 fifteen

25 25

35 35

45 Four. Five

55 55

65 65

de conejo anti-IgG de ratón con los dos conjugados (“amplificación”), como anticuerpo secundario reactivo para detectar la unión a los antígenos tisulares de los anticuerpos primarios abajo relacionados (que suministra Ventana Medical Systems, Inc, Tucson, AZ). Con estos conjugados se trataron cortes tisulares apropiados de archivo y se desarrollaron siguiendo protocolos estándar para la generación de señales detectables mediante HRP (añadiendo DAB) en un aparato Bench-Mark® XT autostainer (Ventana Medical Systems, Inc, Tucson, AZ). Un típico protocolo automatizado incluye la desparafinación, varias etapas de lavado, la adición de un tampón de reacción, la adición del anticuerpo primario, la adición del anticuerpo secundario, la adición de DAB y peróxido de hidrógeno y una tinción de contraste. of rabbit anti-mouse IgG with the two conjugates ("amplification"), as a reactive secondary antibody to detect the binding to the tissue antigens of the primary antibodies listed below (provided by Ventana Medical Systems, Inc, Tucson, AZ). With these conjugates, appropriate tissue cut-offs were treated and developed following standard protocols for the generation of signals detectable by HRP (adding DAB) in a Bench-Mark® XT autostainer (Ventana Medical Systems, Inc, Tucson, AZ). A typical automated protocol includes dewaxing, several washing steps, the addition of a reaction buffer, the addition of the primary antibody, the addition of the secondary antibody, the addition of DAB and hydrogen peroxide and a contrast staining.

Se tiñeron cortes tisulares comparables (adyacentes) con los conjugados revelados y con conjugados de HRP/ F(ab’)2 con andamiaje de polilisina (en lo sucesivo “conjugados con andamiaje”) usados como anticuerpo secundario reactivo. Los conjugados con andamiaje eran de segunda generación (más pequeños y homogéneos según la determinación por cromatografía de exclusión de tamaños) o de primera generación (más grandes e inhomogéneos según la determinación por cromatografía de exclusión de tamaños). Para una descripción más completa de los conjugados con andamiaje véanse las patentes U.S. nº 6,613,564 y 6,252,053. Comparable (adjacent) tissue sections were stained with the disclosed conjugates and HRP / F (ab ’) 2 conjugates with polylysine scaffolding (hereinafter" scaffold conjugates ") used as reactive secondary antibody. The conjugates with scaffolding were second generation (smaller and homogeneous according to the determination by size exclusion chromatography) or first generation (larger and inhomogeneous according to the determination by size exclusion chromatography). For a more complete description of the conjugates with scaffolding see U.S. Pat. 6,613,564 and 6,252,053.

Anticuerpos Antibodies

Anti-bcl-2 (clon 100/D5) Anti-CD15 (clon MMA) Anti-CD20 (clon L26) Anti-PR (clon 16) Anti-EGFR (clon 31G7) Anti-c-erbB-2 (clon CB11) Anti-bcl-2 (clone 100 / D5) Anti-CD15 (clone MMA) Anti-CD20 (clone L26) Anti-PR (clone 16) Anti-EGFR (clone 31G7) Anti-c-erbB-2 (clone CB11)
Anti-CD57 (clon NK-1) Anti-CD23 (clon 1B12) Anti-ER (clon 6F 11) Anti- p53 (clon D07) Anti-ciclina-d1 (clon P2D11F11) Anti-PSA Anti-CD57 (clone NK-1) Anti-CD23 (clone 1B12) Anti-ER (clone 6F 11) Anti-p53 (clone D07) Anti-cyclin-d1 (clone P2D11F11) Anti-PSA

*nota: todos eran anticuerpos de ratón, excepto el PSA, que es un anticuerpo de conejo. * note: all were mouse antibodies, except PSA, which is a rabbit antibody.

La FIG. 2 muestra los resultados de tinción para la detección de bcl-2 mediante el conjugado revelado (FIG. 2A) y el conjugado con andamiaje de segunda generación (FIG. 2B). Los resultados demuestran que la mayor intensidad de tinción se consigue con el conjugado revelado en cortes tisulares comparables. FIG. 2 shows the staining results for the detection of bcl-2 by the conjugate disclosed (FIG. 2A) and the conjugate with second generation scaffolding (FIG. 2B). The results show that the highest intensity of staining is achieved with the conjugate revealed in comparable tissue sections.

La FIG. 3 muestra los resultados de tinción para la detección de CD-15 mediante el conjugado revelado (FIG. 3A) y el conjugado con andamiaje de segunda generación (FIG. 3B). Los resultados demuestran que la mayor intensidad de tinción se consigue con el conjugado revelado en cortes tisulares comparables. FIG. 3 shows the staining results for the detection of CD-15 by the conjugate disclosed (FIG. 3A) and the conjugate with second generation scaffolding (FIG. 3B). The results show that the highest intensity of staining is achieved with the conjugate revealed in comparable tissue sections.

La FIG. 4 muestra los resultados de tinción para la detección de CD-20 mediante el conjugado revelado (usando amplificación, FIG. 4A) y el conjugado con andamiaje de segunda generación (FIG. 4B). Los resultados demuestran que la mayor intensidad de tinción se consigue con el conjugado revelado en cortes tisulares comparables. FIG. 4 shows the staining results for the detection of CD-20 by the conjugate revealed (using amplification, FIG. 4A) and the conjugate with second generation scaffolding (FIG. 4B). The results show that the highest intensity of staining is achieved with the conjugate revealed in comparable tissue sections.

La FIG. 5 muestra los resultados de tinción para la detección de CD-23 mediante el conjugado revelado (FIG. 5A), el conjugado con andamiaje de segunda generación (FIG. 5B) y el conjugado con andamiaje de primera generación (FIG. 5C). Los resultados demuestran que, en cortes tisulares comparables, el conjugado revelado proporciona una intensidad de tinción mayor que la observada para ambos conjugados con andamiaje. FIG. 5 shows the staining results for the detection of CD-23 by the conjugate disclosed (FIG. 5A), the conjugate with second generation scaffolding (FIG. 5B) and the conjugate with first generation scaffolding (FIG. 5C). The results show that, in comparable tissue sections, the conjugate revealed provides a staining intensity greater than that observed for both conjugates with scaffolding.

La FIG. 6 muestra los resultados de tinción para la detección de CD57 mediante el conjugado revelado (FIG. 6A) y el conjugado con andamiaje de segunda generación (FIG. 6B). Los resultados demuestran que la mayor intensidad de tinción se consigue con el conjugado revelado en cortes tisulares comparables. FIG. 6 shows the staining results for the detection of CD57 by the conjugate disclosed (FIG. 6A) and the conjugate with second generation scaffolding (FIG. 6B). The results show that the highest intensity of staining is achieved with the conjugate revealed in comparable tissue sections.

La FIG. 7 muestra los resultados de tinción para la detección de cerb-B2/CB11 mediante el conjugado revelado (FIG. 7A), el conjugado con andamiaje de segunda generación (FIG. 7B) y el conjugado con andamiaje de primera generación (FIG. 7C). Los resultados demuestran que, en cortes tisulares comparables, el conjugado revelado proporciona una intensidad de tinción mayor que la observada para ambos conjugados con andamiaje. FIG. 7 shows the staining results for the detection of cerb-B2 / CB11 by the conjugate disclosed (FIG. 7A), the conjugate with second generation scaffolding (FIG. 7B) and the conjugate with first generation scaffolding (FIG. 7C) . The results show that, in comparable tissue sections, the conjugate revealed provides a staining intensity greater than that observed for both conjugates with scaffolding.

La FIG. 8 muestra los resultados de tinción para la detección de ciclina-D1 mediante el conjugado revelado (FIG. 8A) y el conjugado con andamiaje de segunda generación (FIG. 8B). Los resultados demuestran que la mayor intensidad de tinción se consigue con el conjugado revelado en cortes tisulares comparables. FIG. 8 shows the staining results for the detection of cyclin-D1 by the conjugate disclosed (FIG. 8A) and the conjugate with second generation scaffolding (FIG. 8B). The results show that the highest intensity of staining is achieved with the conjugate revealed in comparable tissue sections.

La FIG. 9 muestra los resultados de tinción para la detección de EGFR mediante el conjugado revelado (FIG. 9A), el conjugado con andamiaje de segunda generación (FIG. 9B) y el conjugado con andamiaje de primera generación (FIG. 9C). Los resultados demuestran que, en cortes tisulares comparables, el conjugado revelado proporciona una intensidad de tinción mayor que la observada para ambos conjugados con andamiaje. FIG. 9 shows the staining results for the detection of EGFR by the conjugate disclosed (FIG. 9A), the conjugate with second generation scaffolding (FIG. 9B) and the conjugate with first generation scaffolding (FIG. 9C). The results show that, in comparable tissue sections, the conjugate revealed provides a staining intensity greater than that observed for both conjugates with scaffolding.

La FIG. 10 muestra los resultados de tinción para la detección de ER mediante el conjugado revelado (FIG. 10A) y el conjugado con andamiaje de segunda generación (FIG. 10B). Los resultados demuestran que la mayor intensidad de tinción se consigue con el conjugado revelado en cortes tisulares comparables. FIG. 10 shows the staining results for the detection of ER by the conjugate disclosed (FIG. 10A) and the conjugate with second generation scaffolding (FIG. 10B). The results show that the highest intensity of staining is achieved with the conjugate revealed in comparable tissue sections.

14 14

imagen13image13

imagen14image14

5 5

10 10

15 fifteen

20 twenty

25 25

30 30

35 35

40 40

45 Four. Five

I. Preparación de un conjugado de anticuerpo de conejo anti-biotina-HRP-PEG12 y su uso para la hibridación metalográfica enzimática in situ I. Preparation of a rabbit anti-biotin-HRP-PEG12 antibody conjugate and its use for in situ enzymatic metallographic hybridization

HRP-PEG12-maleimida (4): en un vial ámbar de 4 ml se introdujeron 18,4 mg (100 eq.) de éster MAL-dPEG12® NHS (Quanta Biodesign, Powell, OH, peso fórmula = 865,92) y luego 341 ul (8,52 mg, 0,213 mM) de HRP (peroxidasa de rábano picante, Pierce, Rockford, IL) en forma de una solución de 25 mg / ml de fosfato sódico 0,1 M, pH 7,5. Luego el vial se puso en un girador automático, en la oscuridad, a la temperatura ambiente (23 – 25ºC) y la reacción de formación del enlace amida se dejo proceder durante 1 hora. Después se tomó un alícuota de 340 µl para purificar. (La capacidad del bucle de inyección del aparato Akta Purifier empleado fue de 500 µl). Luego se obtuvo HRPPEG12-maleimida pura fraccionando la muestra en un aparato Akta Purifier equipado con una columna Superdex 10/300 eluida con 1,0 ml / min de fosfato sódico 0,1 M, pH 7,5. Las fracciones que contenían HRP (F15-17) se reunieron para dar 1,5 ml de una solución de 4,75 mg / ml de HRP-PEG12-maleimida (83,6% de recuperación) según la medición en un espectrofotómetro UV/VIS, usando el coeficiente de extinción a 280 nm de una solución al 1% de pH 7,5, igual a 6,52. HRP-PEG12-maleimide (4): 18.4 mg (100 eq.) Of MAL-dPEG12® NHS ester (Quanta Biodesign, Powell, OH, formula weight = 865.92) were introduced into a 4 ml amber vial and then 341 ul (8.52 mg, 0.213 mM) of HRP (horseradish peroxidase, Pierce, Rockford, IL) in the form of a solution of 25 mg / ml of 0.1 M sodium phosphate, pH 7.5. Then the vial was placed in an automatic spinner, in the dark, at room temperature (23-25 ° C) and the amide bond formation reaction was allowed to proceed for 1 hour. Then an aliquot of 340 µl was taken to purify. (The injection loop capacity of the Akta Purifier apparatus used was 500 µl). Then, pure HRPPEG12-maleimide was obtained by fractionating the sample in an Akta Purifier apparatus equipped with a 10/300 Superdex column eluted with 1.0 ml / min of 0.1 M sodium phosphate, pH 7.5. Fractions containing HRP (F15-17) were pooled to give 1.5 ml of a 4.75 mg / ml solution of HRP-PEG12-maleimide (83.6% recovery) as measured in a UV spectrophotometer / VIS, using the extinction coefficient at 280 nm of a 1% solution of pH 7.5, equal to 6.52.

Anticuerpo de conejo anti-biotina tiol (5): en un vial ámbar de 4 ml se introdujeron 2,0 ml de una solución de 1,0 mg/ml de anticuerpo de conejo anti-biotina (Bethyl, Montgomery, TX). A esta solución se le añadieron luego 105,2 µl de una solución 500 mM del agente reductor DTT (1,4-ditiotreitol) recién preparada. El vial se puso en un girador automático en la oscuridad y la reacción de reducción del disulfuro se dejo proceder durante 25 minutos. La solución se dividió en dos volúmenes iguales (debido a la limitada capacidad de las columnas de desalinización) y el exceso de DTT se eliminó pasando cada una de las fracciones a través de una columna de desalinización PD-10 eluida con fosfato sódico 0,1 M, EDTA 1,0 mM, pH 6,5. Las fracciones que contenían anticuerpo (F4-5) se reunieron para obtener 4,0 ml de una solución de 0,436 mg / ml de anticuerpo de conejo anti-biotina-SH libre de DTT (87,5% de recuperación) según la medición en un espectrofotómetro UV/VIS de Agilent, usando un coeficiente de extinción a 280 nm de una solución al 1% de pH 6,5, igual a 14. Anti-biotin thiol rabbit antibody (5): 2.0 ml of a 1.0 mg / ml solution of rabbit anti-biotin antibody (Bethyl, Montgomery, TX) was introduced into a 4 ml amber vial. To this solution was then added 105.2 µl of a 500 mM solution of the freshly prepared DTT reducing agent (1,4-dithiothreitol). The vial was placed on an automatic spinner in the dark and the disulfide reduction reaction was allowed to proceed for 25 minutes. The solution was divided into two equal volumes (due to the limited capacity of the desalination columns) and the excess DTT was removed by passing each of the fractions through a PD-10 desalination column eluted with 0.1 sodium phosphate. M, 1.0 mM EDTA, pH 6.5. Fractions containing antibody (F4-5) were pooled to obtain 4.0 ml of a solution of 0.436 mg / ml of DTT-free rabbit anti-biotin-SH antibody (87.5% recovery) as measured in an Agilent UV / VIS spectrophotometer, using an extinction coefficient at 280 nm of a 1% solution of pH 6.5, equal to 14.

Conjugación HRP-anticuerpo (6): al anticuerpo de conejo anti-biotina-IgG-tiol (5) se le añadió un exceso tres veces molar de HRP-PEG12-maleimida (4). Luego la reacción se incubó a temperatura ambiente (23 – 25ºC) por la noche. Después de purificarlo a través de una columna Superdex 200 10/300 GL SE se obtuvieron 875 mg de conjugado con un PM medio de 359 kD. HRP-antibody conjugation (6): a threefold excess of HRP-PEG12-maleimide (4) was added to the rabbit anti-biotin-IgG-thiol antibody (5). Then the reaction was incubated at room temperature (23-25 ° C) overnight. After purification through a Superdex 200 10/300 GL SE column, 875 mg of conjugate was obtained with an average MW of 359 kD.

El procedimiento metalográfico enzimático descrito en el ejemplo G se repitió usando el conjugado de PEG12 antibiotina como anticuerpo primario (es decir sin amplificación) y sorprendentemente se obtuvo una tinción intensa, aunque no se empleó ninguna amplificación. Estos resultados demuestran que el empleo de conectores largos de PEG heterobifuncionales (PEG8 o superior, por ejemplo PEG12 o superior) para preparar los conjugados revelados elimina sorprendentemente la necesidad de esquemas de amplificación para las aplicaciones de IHC y ISH en los cortes tisulares. The enzymatic metallographic procedure described in Example G was repeated using the PEG12 antibiotin conjugate as a primary antibody (i.e. without amplification) and surprisingly intense staining was obtained, although no amplification was employed. These results demonstrate that the use of long heterobifunctional PEG connectors (PEG8 or higher, for example PEG12 or higher) to prepare the disclosed conjugates surprisingly eliminates the need for amplification schemes for IHC and ISH applications in tissue cuts.

J. Síntesis del conector de maleimida/hidrazida PEG (ejemplo de referencia) J. Synthesis of the PEG maleimide / hydrazide connector (reference example)

El esquema 6 muestra un método general de preparación de conectores heterobifuncionales de maleimida/hidrazida PEG. En resumen, un conector de maleimida/éster activo de PEG (como el adquirido de Quanta Biodesign) se hace reaccionar con un derivado de hidrazina protegido y después con ácido, para dar el conector de maleimida/hidrazida PEG. Scheme 6 shows a general method of preparing PEG maleimide / hydrazide heterobifunctional connectors. In summary, a PEG active maleimide / ester linker (such as that purchased from Quanta Biodesign) is reacted with a protected hydrazine derivative and then with acid, to give the PEG maleimide / hydrazide connector.

ácido acid

Esquema 6 17 Scheme 6 17

imagen15image15

Claims (1)

imagen1image 1 imagen2image2
ES06758689.1T 2005-04-28 2006-04-27 Enzymes conjugated to antibodies using a heterobifunctional PEG linker Active ES2609919T3 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US67575905P 2005-04-28 2005-04-28
US675759P 2005-04-28
PCT/US2006/016087 WO2006116628A2 (en) 2005-04-28 2006-04-27 Enzymes conjugated to antiobodies via a peg heterobifuctional linker

Publications (1)

Publication Number Publication Date
ES2609919T3 true ES2609919T3 (en) 2017-04-25

Family

ID=37027498

Family Applications (1)

Application Number Title Priority Date Filing Date
ES06758689.1T Active ES2609919T3 (en) 2005-04-28 2006-04-27 Enzymes conjugated to antibodies using a heterobifunctional PEG linker

Country Status (8)

Country Link
US (4) US20060246523A1 (en)
EP (2) EP3144675A1 (en)
JP (1) JP5628476B2 (en)
AU (1) AU2006239315B2 (en)
CA (1) CA2609702C (en)
DK (1) DK1877101T3 (en)
ES (1) ES2609919T3 (en)
WO (1) WO2006116628A2 (en)

Families Citing this family (133)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2609919T3 (en) 2005-04-28 2017-04-25 Ventana Medical Systems, Inc. Enzymes conjugated to antibodies using a heterobifunctional PEG linker
JP2008541015A (en) * 2005-04-28 2008-11-20 ベンタナ・メデイカル・システムズ・インコーポレーテツド Nanoparticle conjugate
CA2631005C (en) * 2005-11-23 2017-02-28 Ventana Medical Systems, Inc. Molecular conjugate
US20090163698A1 (en) * 2007-05-11 2009-06-25 John Joseph Grigsby Method for Preparing Antibody Conjugates
AU2009205995B2 (en) * 2008-01-18 2014-04-03 Medimmune, Llc Cysteine engineered antibodies for site-specific conjugation
US20100136584A1 (en) * 2008-09-22 2010-06-03 Icb International, Inc. Methods for using antibodies and analogs thereof
WO2010078376A2 (en) 2008-12-30 2010-07-08 Ventana Medical Systems, Inc. Fc-specific polymer-conjugated antibodies and their diagnostic use
AU2010339464B2 (en) 2009-12-31 2014-10-09 Ventana Medical Systems, Inc. Methods for producing uniquely specific nucleic acid probes
US8877505B2 (en) 2010-02-02 2014-11-04 Ventana Medical Systems, Inc. Composition and method for stabilizing fluorescent particles
EP2539355B1 (en) 2010-02-26 2016-10-05 Ventana Medical Systems, Inc. In-situ hybridization with polytag probes
AU2011220792A1 (en) 2010-02-26 2012-07-26 Ventana Medical Systems, Inc. Cytogenic analysis of metaphase chromosomes
CA2936532A1 (en) 2010-04-20 2011-10-27 Ventana Medical Systems, Inc. Two-color chromogenic in situ hybridization
EP2380909A1 (en) 2010-04-26 2011-10-26 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. PTK-7 protein involved in breast cancer
DK2588144T3 (en) 2010-07-02 2018-07-23 Ventana Med Syst Inc Detection of targets using mass markers and mass spectrometry
RU2598711C2 (en) 2010-07-22 2016-09-27 Те Риджентс Оф Те Юниверсити Оф Калифорния Anti-tumour antigen antibody and methods of application
WO2012024185A1 (en) 2010-08-16 2012-02-23 Ventana Medical Systems, Inc. Substrates for chromogenic detection and methods of use in detection assays and kits
EP2638392B1 (en) * 2010-11-08 2016-01-13 Dako Denmark A/S Quantification of single target molecules in histological samples
US10718777B2 (en) 2010-12-06 2020-07-21 Agilent Technologies, Inc. Combined histological stain
CA2825453C (en) 2011-03-14 2016-05-10 Ventana Medical Systems, Inc. A method of analyzing chromosomal translocations and a system therefore
WO2013016449A2 (en) 2011-07-26 2013-01-31 Indicator Systems International, Inc. Assays for the detection of microbes
WO2013167387A1 (en) 2012-05-10 2013-11-14 Ventana Medical Systems, Inc. Uniquely specific probes for pten, pik3ca, met, top2a, and mdm2
CA3062003C (en) 2012-05-17 2022-01-11 Extend Biosciences, Inc. Vitamin d as a targeting group for therapeutic peptides
ES2648176T3 (en) 2012-07-12 2017-12-28 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting survival time and response to treatment of a patient suffering from solid cancer with a hallmark of at least 7 genes
DK2880180T3 (en) 2012-08-06 2019-01-14 Inst Nat Sante Rech Med METHODS AND KITS TO SCREEN PATIENTS WITH CANCER
WO2014048942A1 (en) 2012-09-25 2014-04-03 Ventana Medical Systems, Inc. Probes for pten, pik3ca, met, and top2a, and method for using the probes
US20140134651A1 (en) * 2012-11-07 2014-05-15 Quanta Eqip, Llc Stable Discrete PEG Based Peroxidase Biological Conjugates
CN105849086B (en) 2012-11-24 2018-07-31 杭州多禧生物科技有限公司 Hydrophily chain junctor and its application on drug molecule and cell-binding molecules conjugation reaction
AU2014230945B2 (en) 2013-03-12 2019-07-11 Ventana Medical Systems, Inc. Proximity assay for in situ detection of targets
US9889203B2 (en) * 2013-03-15 2018-02-13 Amicus Therapeutics, Inc. Chemical crosslinkers
WO2015001082A1 (en) 2013-07-05 2015-01-08 INSERM (Institut National de la Santé et de la Recherche Médicale) Novel alternative splice transcripts for mhc class i related chain alpha (mica) and uses thereof
WO2015036405A1 (en) 2013-09-10 2015-03-19 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for diagnosing and treating basal cell carcinoma
CN114262344A (en) 2014-02-28 2022-04-01 杭州多禧生物科技有限公司 Charged linkers and their use in conjugation reactions
CN106415269B (en) * 2014-05-08 2020-11-27 贵州美鑫达医疗科技有限公司 Direct immunohistochemical assay
EP3009147A1 (en) 2014-10-16 2016-04-20 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for treating resistant glioblastoma
US9789197B2 (en) 2014-10-22 2017-10-17 Extend Biosciences, Inc. RNAi vitamin D conjugates
EP3220961B1 (en) 2014-10-22 2023-07-05 Extend Biosciences, Inc. Therapeutic vitamin d conjugates
WO2016065052A1 (en) 2014-10-22 2016-04-28 Extend Biosciences, Inc. Insulin vitamin d conjugates
EP3224621B1 (en) 2014-11-25 2019-06-12 Ventana Medical Systems, Inc. Proximity assays using chemical ligation and hapten transfer
EP4170346A1 (en) 2015-02-20 2023-04-26 Idexx Laboratories, Inc. Homogenous immunoassay with compensation for background signal
CN106338423B (en) 2015-07-10 2020-07-14 三斯坎公司 Spatial multiplexing of histological staining
CN108369229A (en) * 2015-07-17 2018-08-03 奥菲迪亚有限公司 Connection molecule for handling substrate surface
JP6675165B2 (en) 2015-07-31 2020-04-01 シスメックス株式会社 Test substance detection method, detection reagent kit and detection reagent
WO2017029391A1 (en) 2015-08-20 2017-02-23 INSERM (Institut National de la Santé et de la Recherche Médicale) New method for treating cancer
EP3341734B1 (en) 2015-08-28 2024-08-07 Ventana Medical Systems, Inc. Protein proximity assay in formalin fixed paffafin embedded tissue using caged haptens
WO2017055327A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of endothelial cells in a tissue sample
WO2017055322A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of neutrophils in a tissue sample
WO2017055326A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of myeloid dendritic cells in a tissue sample
WO2017055321A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of fibroblasts in a tissue sample
WO2017055324A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of cells of monocytic origin in a tissue sample
WO2017055319A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of b cells in a tissue sample
WO2017055320A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of cytotoxic lymphocytes in a tissue sample
WO2017055325A1 (en) 2015-09-29 2017-04-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of nk cells in a tissue sample
WO2017060397A1 (en) 2015-10-09 2017-04-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting the survival time of subjects suffering from melanoma metastases
WO2017067944A1 (en) 2015-10-19 2017-04-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting the survival time of subjects suffering from triple negative breast cancer
CA3001727A1 (en) * 2015-11-18 2017-05-26 Synergy Pharmaceuticals, Inc. Compositions and method for the treatment and detection of colon cancer
WO2017155996A1 (en) 2016-03-08 2017-09-14 Ventana Medical Systems, Inc. Multiplexed immunohistochemistry using recombinant antibodies with epitope tags
US10900069B2 (en) 2016-03-10 2021-01-26 Emory University Devices and methods useful for detecting mechanical forces of ligand receptor interactions
WO2017182834A1 (en) 2016-04-19 2017-10-26 INSERM (Institut National de la Santé et de la Recherche Médicale) New method for treating resistant glioblastoma
EP3463452A1 (en) 2016-05-24 2019-04-10 Institut National de la Sante et de la Recherche Medicale (INSERM) Methods and pharmaceutical compositions for the treatment of non small cell lung cancer (nsclc) that coexists with chronic obstructive pulmonary disease (copd)
CN116840465A (en) 2016-06-28 2023-10-03 文塔纳医疗系统公司 New colors dyed with chromogenic IHC and ISH of multi-dye quinone methide and tyramide conjugates
WO2018002016A1 (en) 2016-06-28 2018-01-04 Ventana Medical Systems, Inc. Application of click chemistry for signal amplification in ihc and ish assays
WO2018011107A1 (en) 2016-07-11 2018-01-18 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of er-alpha 46 in methods and kits for assessing the status of breast cancer
WO2018011166A2 (en) 2016-07-12 2018-01-18 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for quantifying the population of myeloid dendritic cells in a tissue sample
CN106405087A (en) * 2016-08-31 2017-02-15 上海科华生物工程股份有限公司 Goat anti-HBsAg polyclonal antibody-alkaline phosphatase conjugate, and preparation method and detection kit thereof
WO2018046736A1 (en) 2016-09-12 2018-03-15 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting the survival time of patients suffering from cancer
EP3515453A1 (en) 2016-09-22 2019-07-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for reprograming immune environment in a subject in need thereof
US11525008B2 (en) 2016-09-22 2022-12-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for the treatment of lung cancer
WO2018122249A1 (en) 2016-12-28 2018-07-05 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting the survival time of patients suffering from a microsatellite stable colorectal cancer
WO2018122245A1 (en) 2016-12-28 2018-07-05 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods of predicting the survival time of patients suffering from cms3 colorectal cancer
WO2018146239A1 (en) 2017-02-10 2018-08-16 INSERM (Institut National de la Santé et de la Recherche Médicale) Biomarker for outcome in aml patients
WO2018162404A1 (en) 2017-03-06 2018-09-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Biomarker for outcome in aml patients
WO2018172540A1 (en) 2017-03-24 2018-09-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Method to predict the progression of alzheimer's disease
WO2018189215A1 (en) 2017-04-12 2018-10-18 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for predicting the survival time of a patient suffering from hepatocellular carcinoma
GB201709503D0 (en) * 2017-06-15 2017-08-02 Method and device for assay improvement
WO2018232088A1 (en) * 2017-06-16 2018-12-20 Eli Lilly And Company Engineered antibody compounds and conjugates thereof
WO2019038219A1 (en) 2017-08-21 2019-02-28 INSERM (Institut National de la Santé et de la Recherche Médicale) New prognostic method of pancreatic cancer
WO2019043138A1 (en) 2017-09-01 2019-03-07 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for predicting the outcome of a cancer
AU2018351349A1 (en) * 2017-10-19 2020-04-23 Idexx Laboratories, Inc. Detection of symmetrical dimethylarginine
JP7038209B2 (en) 2017-11-13 2022-03-17 エフ．ホフマン－ラ ロシュ アーゲー Equipment for sample analysis using epitaco electrophoresis
WO2019207030A1 (en) 2018-04-26 2019-10-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting a response with an immune checkpoint inhibitor in a patient suffering from a lung cancer
WO2020074742A1 (en) 2018-10-12 2020-04-16 F. Hoffmann-La Roche Ag Detection methods for epitachophoresis workflow automation
WO2020089432A1 (en) 2018-11-02 2020-05-07 INSERM (Institut National de la Santé et de la Recherche Médicale) New prognostic method of pancreatic cancer
WO2020089428A1 (en) 2018-11-02 2020-05-07 INSERM (Institut National de la Santé et de la Recherche Médicale) New prognostic method of pancreatic cancer
CA3125476A1 (en) 2019-01-03 2020-07-09 Inserm (Institut National De La Sante Et De La Recherche Medicale) Methods and pharmaceutical compositions for enhancing cd8+ t cell-dependent immune responses in subjects suffering from cancer
EP3911669A1 (en) 2019-01-16 2021-11-24 INSERM (Institut National de la Santé et de la Recherche Médicale) Variants of erythroferrone and their use
WO2020165370A1 (en) 2019-02-13 2020-08-20 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and compositions for selecting a cancer treatment in a subject suffering from cancer
WO2020182932A1 (en) 2019-03-13 2020-09-17 INSERM (Institut National de la Santé et de la Recherche Médicale) New gene signatures for predicting survival time in patients suffering from renal cell carcinoma
WO2020193740A1 (en) 2019-03-28 2020-10-01 INSERM (Institut National de la Santé et de la Recherche Médicale) New strategy for treating pancreatic cancer
WO2020201362A2 (en) 2019-04-02 2020-10-08 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods of predicting and preventing cancer in patients having premalignant lesions
EP3959340A1 (en) 2019-04-24 2022-03-02 Institut National de la Santé et de la Recherche Médicale (INSERM) Method for predicting the response of antipsychotic drugs
WO2020229521A1 (en) 2019-05-14 2020-11-19 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for inhibiting or reducing bacterial biofilms on a surface
US20220325268A1 (en) 2019-05-14 2022-10-13 Roche Sequencing Solutions, Inc Devices and methods for sample analysis
CN113785184B (en) 2019-05-14 2025-02-28 文塔纳医疗系统公司 System including a biological sample processing chamber
AU2020288603A1 (en) 2019-06-03 2021-12-16 Assistance Publique - Hôpitaux De Paris Methods for modulating a treatment regimen
WO2021001539A1 (en) 2019-07-04 2021-01-07 INSERM (Institut National de la Santé et de la Recherche Médicale) New strategy to detect and treat eosinophilic fasciitis
WO2021044012A1 (en) 2019-09-05 2021-03-11 INSERM (Institut National de la Santé et de la Recherche Médicale) Method of treatment and pronostic of acute myeloid leukemia
CA3157889A1 (en) 2019-10-17 2021-04-22 Inserm (Institut National De La Sante Et De La Recherche Medicale) Methods for diagnosing nasal intestinal type adenocarcinomas
JP7649623B2 (en) 2019-12-23 2025-03-21 株式会社Ｇｓユアサ Method for estimating SOC of a valve-regulated lead-acid battery and device for monitoring the valve-regulated lead-acid battery
WO2021170777A1 (en) 2020-02-28 2021-09-02 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for diagnosing, prognosing and managing treatment of breast cancer
EP4121768A1 (en) 2020-03-20 2023-01-25 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for predicting the survival time of a patient suffering from a cancer
JP7515855B2 (en) * 2020-04-13 2024-07-16 国立研究開発法人産業技術総合研究所 Sugar chain ligand with high effect of suppressing non-specific adsorption, and toxin detection chip having said sugar chain ligand immobilized thereon
US20230250426A1 (en) 2020-06-10 2023-08-10 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for treating and prognosing cancer like glioblastoma
EP4168006A1 (en) 2020-06-18 2023-04-26 Institut National de la Santé et de la Recherche Médicale (INSERM) New strategy for treating pancreatic cancer
CN115997123A (en) 2020-06-30 2023-04-21 国家医疗保健研究所 Methods for predicting risk of recurrence and/or death of solid cancer patients after preoperative adjuvant therapy
EP4172628A1 (en) 2020-06-30 2023-05-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting the risk of recurrence and/or death of patients suffering from a solid cancer after preoperative adjuvant therapy and radical surgery
WO2022018163A1 (en) 2020-07-22 2022-01-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for predicting survival time in patients suffering from cancer
WO2022064049A1 (en) 2020-09-28 2022-03-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for diagnosing brucella infection
WO2022084327A1 (en) 2020-10-20 2022-04-28 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for predicting the response to tnf inhibitors
CN112305222A (en) * 2020-11-05 2021-02-02 东莞市医本生物科技有限公司 Small polymer enzyme-antibody fragment, preparation and application thereof
WO2022096633A1 (en) 2020-11-06 2022-05-12 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for diagnosis and treating polycystic ovary syndrome (pcos)
WO2022135753A1 (en) 2020-12-21 2022-06-30 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for prognosis the humoral response of a subject prior to vaccination
WO2022136252A1 (en) 2020-12-21 2022-06-30 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for prognosis the humoral response of a subject prior to vaccination
WO2022152698A1 (en) 2021-01-12 2022-07-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of npdk-d to evaluate cancer prognosis
WO2022152777A1 (en) 2021-01-15 2022-07-21 F. Hoffmann-La Roche Ag Storage stable caged haptens
CN116761819A (en) * 2021-01-18 2023-09-15 味之素株式会社 Compounds or salts thereof, and antibodies derived therefrom
JP2024507677A (en) 2021-01-25 2024-02-21 ベンタナ メディカル システムズ， インコーポレイテッド A stained biological specimen containing one or more biomarkers labeled with one or more detectable moieties
EP4291897A1 (en) 2021-02-09 2023-12-20 Institut National de la Santé et de la Recherche Médicale (INSERM) New method to pronostic lung cancer
EP4308934A1 (en) 2021-03-17 2024-01-24 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for diagnosing pancreatic cancer
WO2022207566A1 (en) 2021-03-29 2022-10-06 INSERM (Institut National de la Santé et de la Recherche Médicale) New method to evaluate pancreatic cancer prognosis
WO2022223791A1 (en) 2021-04-23 2022-10-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and compositions for treating cell senescence accumulation related disease
EP4367269A1 (en) 2021-07-05 2024-05-15 Inserm (Institut National De La Sante Et De La Recherche Medicale) Gene signatures for predicting survival time in patients suffering from renal cell carcinoma
WO2023089159A1 (en) 2021-11-22 2023-05-25 INSERM (Institut National de la Santé et de la Recherche Médicale) New strategy targeting stroma/tumor cell crosstalk to treat a cancer
EP4472740A1 (en) 2022-01-31 2024-12-11 INSERM (Institut National de la Santé et de la Recherche Médicale) Cd38 as a biomarker and biotarget in t-cell lymphomas
WO2023152133A1 (en) 2022-02-08 2023-08-17 INSERM (Institut National de la Santé et de la Recherche Médicale) Method for diagnosing colorectal cancer
AU2022447608A1 (en) 2022-03-17 2024-09-26 Assistance Publique - Hôpitaux De Paris Methods for predicting response to an immunotherapeutic treatment in a patient with a cancer
US20230408497A1 (en) * 2022-06-20 2023-12-21 Sysmex Corporation Labeled polypeptide, modified polypeptide, production method for these polypeptides, reagent containing these polypeptides, and measurement method for target substance
WO2024061930A1 (en) 2022-09-22 2024-03-28 Institut National de la Santé et de la Recherche Médicale New method to treat and diagnose peripheral t-cell lymphoma (ptcl)
US12233115B2 (en) 2022-09-30 2025-02-25 Extend Biosciences, Inc. Long-acting parathyroid hormone
WO2024115935A1 (en) 2022-11-29 2024-06-06 Inserm Methods for the treatment of b-cell lymphoma using cd39 inhibitors
WO2024236131A1 (en) 2023-05-17 2024-11-21 Institut National de la Santé et de la Recherche Médicale Stratificate and method to treat a patient suffering from a cancer
WO2024245951A1 (en) 2023-05-26 2024-12-05 Institut National de la Santé et de la Recherche Médicale Combination of slc8a1 inhibitor and mitochondria-targeted antioxidant for treating melanoma
WO2025027127A1 (en) 2023-08-02 2025-02-06 Institut National de la Santé et de la Recherche Médicale New prognostic method of kidney failure
CN118373870B (en) * 2024-05-10 2024-11-26 中汉生物医学技术研究(昆山)有限公司 A protein coupling method based on aldehyde-thiol reaction

Family Cites Families (70)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB782420A (en) 1954-08-23 1957-09-04 Ici Ltd New hydrazides
US3654090A (en) * 1968-09-24 1972-04-04 Organon Method for the determination of antigens and antibodies
NL154599B (en) * 1970-12-28 1977-09-15 Organon Nv PROCEDURE FOR DETERMINING AND DETERMINING SPECIFIC BINDING PROTEINS AND THEIR CORRESPONDING BINDABLE SUBSTANCES, AND TEST PACKAGING.
US4002532A (en) * 1974-10-21 1977-01-11 Weltman Joel K Enzyme conjugates
IL47468A (en) * 1975-06-12 1979-05-31 Rehovot Res Prod Process for the cross-linking of proteins using water soluble cross-linking agents
US4016043A (en) * 1975-09-04 1977-04-05 Akzona Incorporated Enzymatic immunological method for the determination of antigens and antibodies
JPS5344622A (en) * 1976-09-30 1978-04-21 Mochida Pharm Co Ltd Immunologically measuring method
DE2708018A1 (en) * 1977-02-24 1978-09-07 Boehringer Mannheim Gmbh BIOLOGICALLY ACTIVE PROTEIN FIXED TO POLYAMIDE AND THE PROCESS FOR ITS PRODUCTION
SE427505B (en) * 1977-03-04 1983-04-11 Pharmacia Diagnostics Ab REAGENT USE FOR IMMUNKEMIC DETERMINATION METHODS
US4235960A (en) * 1977-07-29 1980-11-25 The Medical College Of Wisconsin, Inc. Competitive enzyme-linked immunoassay
US4218539A (en) * 1978-03-24 1980-08-19 Weltman Joel K Enzyme conjugates and method of preparation and use
US4433059A (en) * 1981-09-08 1984-02-21 Ortho Diagnostic Systems Inc. Double antibody conjugate
US4671958A (en) * 1982-03-09 1987-06-09 Cytogen Corporation Antibody conjugates for the delivery of compounds to target sites
US4454226A (en) * 1982-03-17 1984-06-12 Majid Ali Enzymatic immunoassay
US4657863A (en) 1982-07-02 1987-04-14 Celanese Corporation Stabilization of a mutant microorganism population
US4657853A (en) * 1984-09-14 1987-04-14 E. I. Du Pont De Nemours And Company Immunoassays utilizing covalent conjugates of polymerized enzyme and antibody
US4732863A (en) * 1984-12-31 1988-03-22 University Of New Mexico PEG-modified antibody with reduced affinity for cell surface Fc receptors
US5057313A (en) * 1986-02-25 1991-10-15 The Center For Molecular Medicine And Immunology Diagnostic and therapeutic antibody conjugates
US4810638A (en) 1986-07-24 1989-03-07 Miles Inc. Enzyme-labeled antibody reagent with polyalkyleneglycol linking group
US5002883A (en) * 1987-10-30 1991-03-26 Abbott Laboratories Covalent attachment of antibodies and antigens to solid phases using extended length heterobifunctional coupling agents
US4994385A (en) * 1987-10-30 1991-02-19 Abbott Laboratories Heterobifunctional coupling agents
US5053520A (en) 1988-09-22 1991-10-01 Abbott Laboratories Heterobifunctional maleimido containing coupling agents
US5063109A (en) * 1988-10-11 1991-11-05 Abbott Laboratories Covalent attachment of antibodies and antigens to solid phases using extended length heterobifunctional coupling agents
US5157123A (en) 1989-03-13 1992-10-20 Georgetown University S-(2-thiopyridyl)-l-cysteine, a heterobifunctional crosslinking reagent
DK0555336T3 (en) 1990-10-22 1996-11-25 Abbott Lab Homobifunctional agents for coupling enzymes and the like to antibodies and the like
US5191066A (en) * 1990-12-07 1993-03-02 Abbott Laboratories Site-specific conjugation of immunoglobulins and detectable labels
US6787153B1 (en) 1991-06-28 2004-09-07 Mitsubishi Chemical Corporation Human monoclonal antibody specifically binding to surface antigen of cancer cell membrane
JP3236667B2 (en) * 1991-06-28 2001-12-10 三菱化学株式会社 Human monoclonal antibody and gene encoding the same, hybridoma and antitumor agent
US5329028A (en) * 1992-08-05 1994-07-12 Genentech, Inc. Carbohydrate-directed cross-linking reagents
US6005079A (en) 1992-08-21 1999-12-21 Vrije Universiteit Brussels Immunoglobulins devoid of light chains
EP0656946B2 (en) 1992-08-21 2010-03-31 Vrije Universiteit Brussel Immunoglobulins devoid of light chains
US5648218A (en) * 1993-02-12 1997-07-15 Sealite Sciences, Inc. Preparation of photoprotein conjugates and methods of use thereof
US5736624A (en) * 1994-12-02 1998-04-07 Abbott Laboratories Phosphatase activated crosslinking, conjugating and reducing agents; methods of using such agents; and reagents comprising phosphatase activated crosslinking and conjugating agents
DE59607898D1 (en) * 1995-12-29 2001-11-15 Biotez Berlin Buch Gmbh Bioche METHOD FOR MARKING BIOMOLECULES WITH HORSERADISH PEROXIDASE
KR19990029749A (en) * 1997-09-17 1999-04-26 미우라 아끼라 Divalent reactive water soluble polymer derivatives and composites containing them
JP4385152B2 (en) * 1997-09-17 2009-12-16 三菱化学株式会社 Bivalent reactive water-soluble polymer derivative and complex containing the same
US6218160B1 (en) * 1997-10-31 2001-04-17 Roche Diagnostics Corporation Site-specific conjugation of glycoproteins
JP3524401B2 (en) 1998-09-16 2004-05-10 株式会社ニチレイ Enzyme-antibody complex and method for producing the same
ATE234468T1 (en) 1998-09-18 2003-03-15 Massachusetts Inst Technology BIOLOGICAL USES OF SEMICONDUCTING NANOCRYSTALS
US6576746B2 (en) * 1998-10-13 2003-06-10 Immunomedics, Inc. Site-specific labeling of disulfide-containing targeting vectors
AU3191100A (en) * 1999-03-17 2000-10-04 Mitsubishi Chemical Corporation Ligand-bonded complex
US20050074499A1 (en) * 1999-03-17 2005-04-07 Mitsubishi Chemical Corporation Ligand-bonded complex
ATE439452T1 (en) * 1999-05-07 2009-08-15 Life Technologies Corp METHOD FOR DETECTING ANALYTES USING SEMICONDUCTOR NANOCRYSTALS
GB9919338D0 (en) * 1999-08-16 1999-10-20 Celltech Therapeutics Ltd Biological products
JP3781934B2 (en) * 1999-12-22 2006-06-07 株式会社ニチレイバイオサイエンス Enzyme-protein complex
EP1118334A1 (en) * 2000-01-11 2001-07-25 Aventis Behring Gesellschaft mit beschränkter Haftung Method for the production of conjugates and uses thereof for the prevention and treatment of allergic reactions and autoimmune diseases
EP1118335A1 (en) * 2000-01-11 2001-07-25 Aventis Behring GmbH Method for the production of conjugates for the treatment of allergic reactions and autoimmune diseases
EP2298736B1 (en) * 2000-03-22 2015-07-15 Solulink, Inc. Hydrazine-based and carbonyl-based bifunctional crosslinking reagents
US6649138B2 (en) * 2000-10-13 2003-11-18 Quantum Dot Corporation Surface-modified semiconductive and metallic nanoparticles having enhanced dispersibility in aqueous media
US20020083888A1 (en) * 2000-12-28 2002-07-04 Zehnder Donald A. Flow synthesis of quantum dot nanocrystals
US6670113B2 (en) * 2001-03-30 2003-12-30 Nanoprobes Enzymatic deposition and alteration of metals
EP2218762A3 (en) * 2001-07-20 2010-09-29 Life Technologies Corporation Luminescent nanoparticles and methods for their preparation
CA2462930C (en) 2001-10-10 2012-07-10 Shawn De Frees Remodeling and glycoconjugation of peptides
US20030149246A1 (en) * 2002-02-01 2003-08-07 Russell John C. Macromolecular conjugates and processes for preparing the same
US20060153839A1 (en) * 2002-09-16 2006-07-13 Elusys Therapeutics, Inc. Production of bispecific molecules using polyethylene glycol linkers
US7217845B2 (en) * 2002-11-25 2007-05-15 Sun Bio, Inc. Bifunctional polyethylene glycol derivatives
US7888536B2 (en) * 2004-02-13 2011-02-15 Quanta Biodesign, Ltd. Selective and specific preparation of discrete PEG compounds
US20050176108A1 (en) * 2003-03-13 2005-08-11 Young-Min Kim Physiologically active polypeptide conjugate having prolonged in vivo half-life
EP2530719B1 (en) 2003-05-07 2020-08-05 Indiana University Research and Technology Corporation Alloyed semiconductor concentration-gradient quantum dots, use and method of fabricating thereof
EP1636586B1 (en) * 2003-06-24 2009-07-22 Ventana Medical Systems, Inc. Enzyme-catalyzed metal deposition for the enhanced in situ detection of immunohistochemical epitopes and nucleic acid sequences
US7642064B2 (en) * 2003-06-24 2010-01-05 Ventana Medical Systems, Inc. Enzyme-catalyzed metal deposition for the enhanced detection of analytes of interest
KR100657891B1 (en) * 2003-07-19 2006-12-14 삼성전자주식회사 Semiconductor Nanocrystals and Manufacturing Method Thereof
WO2005064018A2 (en) * 2003-12-22 2005-07-14 Ventana Medical Systems, Inc. Microwave mediated synthesis of nucleic acid probes
US20050186642A1 (en) * 2004-02-24 2005-08-25 Biocare Medical, Inc. Immunoassay reagents and methods of use thereof
GB0412181D0 (en) * 2004-06-01 2004-06-30 Celltech R&D Ltd Biological products
US7361516B2 (en) 2004-09-24 2008-04-22 The United States Of America As Represented By The Secretary Of The Navy Field of modular multifunctional ligands
JP2008541015A (en) * 2005-04-28 2008-11-20 ベンタナ・メデイカル・システムズ・インコーポレーテツド Nanoparticle conjugate
ES2609919T3 (en) * 2005-04-28 2017-04-25 Ventana Medical Systems, Inc. Enzymes conjugated to antibodies using a heterobifunctional PEG linker
US20070122408A1 (en) * 2005-10-20 2007-05-31 The Scripps Research Institute Fc Labeling for Immunostaining and Immunotargeting
CA2631005C (en) * 2005-11-23 2017-02-28 Ventana Medical Systems, Inc. Molecular conjugate

Also Published As

Publication number Publication date
EP1877101B1 (en) 2016-11-16
AU2006239315B2 (en) 2012-03-01
CA2609702A1 (en) 2006-11-02
US20160187324A1 (en) 2016-06-30
JP5628476B2 (en) 2014-11-19
EP3144675A1 (en) 2017-03-22
CA2609702C (en) 2013-05-28
US20140147906A1 (en) 2014-05-29
DK1877101T3 (en) 2017-01-09
US20060246523A1 (en) 2006-11-02
WO2006116628A3 (en) 2007-12-13
US8658389B2 (en) 2014-02-25
US11359185B2 (en) 2022-06-14
US20090176253A1 (en) 2009-07-09
US9315789B2 (en) 2016-04-19
WO2006116628A2 (en) 2006-11-02
JP2008539270A (en) 2008-11-13
EP1877101A2 (en) 2008-01-16
AU2006239315A1 (en) 2006-11-02

Similar Documents

Publication Publication Date Title
ES2609919T3 (en) Enzymes conjugated to antibodies using a heterobifunctional PEG linker
Shabat Self‐immolative dendrimers as novel drug delivery platforms
Versteegen et al. Click to release: instantaneous doxorubicin elimination upon tetrazine ligation
de Groot et al. “Cascade‐release dendrimers” liberate all end groups upon a single triggering event in the dendritic core
Hammann et al. Folding and activity of the hammerhead ribozyme
JP2509795B2 (en) Endogenous factor-horseradish radish peroxidase conjugates and methods to increase their stability
WO2005037992A3 (en) Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates, and methods of making said conjugates
Schneider et al. Dextramabs: a novel format of antibody‐drug conjugates featuring a multivalent polysaccharide scaffold
Rodrigues et al. Phosphate-mediated coanchoring of RBD immunogens and molecular adjuvants to alum potentiates humoral immunity against SARS-CoV-2
Jbara et al. Oligonucleotide bioconjugation with bifunctional palladium reagents
Sunasee et al. Covalent and noncovalent bioconjugation strategies
Lai et al. Enhanced cytotoxicity of saporin by polyamidoamine dendrimer conjugation and photochemical internalization
Shabat et al. Chemical adaptor systems
Refsnes et al. Concentration‐dependent cytokine responses of silica nanoparticles and role of ROS in human lung epithelial cells
JP2011173960A (en) Polymer micelle type photostimulation-responsive nitrogen monoxide donor
Park et al. Excited-state tautomerization dynamics of 7-hydroxyquinoline in β-cyclodextrin
Zhang et al. Switchable Enzymatic Accessibility for Precision Cell‐Selective Surface Glycan Remodeling
Trattnig et al. Site‐specific multi‐functionalization of the carrier protein CRM197 by disulfide rebridging for conjugate vaccine development
HUP0100350A2 (en) Conjugates useful in the treatment of prostate cancer, pharmaceutical compositions comprising thereof, process for their preparation and their use
Reille‐Seroussi et al. Genetic encoding and enzymatic deprotection of a latent thiol side chain to enable new protein bioconjugation applications
US8153111B2 (en) Photo-triggered release of active substances from dendrimer-photosensitizer complexes
Gray et al. A comparison of the activity, lysosomal stability, and efficacy of legumain-cleavable and cathepsin-cleavable ADC linkers
EP1161685B1 (en) Method for reducing fluorescence quenching in bioassays
Metelev et al. Synthesis and properties of fluorescent NF-κB-recognizing hairpin oligodeoxyribonucleotide decoys
Agafonov et al. C‐terminal modifications of a protein by UAG‐encoded incorporation of puromycin during in vitro protein synthesis in the absence of release factor 1